A simple in vitro assay for assessing the efficacy, mechanisms and kinetics of anti-prion fibril compounds

Prion
Carol L Ladner-KeayDavid S Wishart

Abstract

Prion diseases are caused by the conversion of normal cellular prion proteins (PrP) into lethal prion aggregates. These prion aggregates are composed of proteinase K (PK) resistant fibrils and comparatively PK-sensitive oligomers. Currently there are no anti-prion pharmaceuticals available to treat or prevent prion disease. Methods of discovering anti-prion molecules rely primarily on relatively complex cell-based, tissue slice or animal-model assays that measure the effects of small molecules on the formation of PK-resistant prion fibrils. These assays are difficult to perform and do not detect the compounds that directly inhibit oligomer formation or alter prion conversion kinetics. We have developed a simple cell-free method to characterize the impact of anti-prion fibril compounds on both the oligomer and fibril formation. In particular, this assay uses shaking-induced conversion (ShIC) of recombinant PrP in a 96-well format and resolution enhanced native acidic gel electrophoresis (RENAGE) to generate, assess and detect PrP fibrils in a high throughput fashion. The end-point PrP fibrils from this assay can be further characterized by PK analysis and negative stain transmission electron microscopy (TEM). This cell-free, gel...Continue Reading

References

Aug 1, 1992·Journal of Neurochemistry·B Caughey, R E Race
Jan 1, 1995·Journal of Virology·L IngrossoM Pocchiari
Apr 3, 2001·Annual Review of Neuroscience·J Collinge
Mar 26, 2002·The Journal of Biological Chemistry·Ilia V BaskakovFred E Cohen
Aug 1, 2002·Proceedings of the National Academy of Sciences of the United States of America·Gianluigi ForloniFabrizio Tagliavini
Sep 27, 2002·Annals of Neurology·Steven J CollinsColin L Masters
Apr 15, 2003·Journal of Virology·Byron CaugheyGerald S Baron
Jul 15, 2003·Journal of Virology·A BarretJ P Deslys
Sep 13, 2005·Biochemical and Biophysical Research Communications·Kenjiro OnoMasahito Yamada
Sep 20, 2005·The Journal of Nutritional Biochemistry·Jong-Deog KimMohsen Meydani
Jul 10, 2007·Proceedings of the National Academy of Sciences of the United States of America·Kazuo KuwataShigeru Katamine
Nov 10, 2007·Science·John Collinge, Anthony R Clarke
Jan 17, 2008·The Journal of General Virology·Constanze RiemerMichael Baier
Jan 29, 2008·Nature Chemical Biology·Brian Y FengBrian K Shoichet
Mar 22, 2008·European Journal of Neurology : the Official Journal of the European Federation of Neurological Societies·I BoneJ Darbyshire
Dec 2, 2008·Neurochemistry International·Saravanan S KaruppagounderGary E Gibson
Dec 17, 2008·Proceedings of the National Academy of Sciences of the United States of America·Christina J SigurdsonAdriano Aguzzi
Sep 2, 2009·Journal of Molecular Biology·Maurizio PolanoGiuseppe Legname
Oct 1, 2009·Neuropathology : Official Journal of the Japanese Society of Neuropathology·Yoshio TsuboiTatsuo Yamada
Nov 7, 2009·The American Journal of Pathology·Tsuyoshi HamaguchiMasahito Yamada
Nov 17, 2009·Proceedings of the National Academy of Sciences of the United States of America·David W ColbyStanley B Prusiner
Dec 25, 2009·Journal of Virology·Sina GhaemmaghamiStanley B Prusiner
Jun 1, 2010·The Journal of Biological Chemistry·Ali Reza A LadiwalaPeter M Tessier
Jan 8, 2011·The Journal of Biological Chemistry·Zaira E Arellano AnayaDidier Vilette
May 4, 2011·Chembiochem : a European Journal of Chemical Biology·Alaina S DeTomaMi Hee Lim
May 26, 2011·The Journal of Biological Chemistry·Guillaume Poncet-MontangeSina Ghaemmaghami
Mar 7, 2012·The Journal of Biological Chemistry·Kenjiro OnoMasahito Yamada
Aug 24, 2013·The Journal of Pharmacology and Experimental Therapeutics·Duo LuStanley B Prusiner
Oct 15, 2013·Neurology·Michael D GeschwindBruce L Miller
Jan 1, 2014·Structure·Pravas Kumar BaralMichael N G James
Apr 26, 2014·Journal of Medicinal Chemistry·Sina GhaemmaghamiAdam R Renslo
May 14, 2014·Prion·Fozia SaleemDavid S Wishart

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Citations

Jul 5, 2019·Expert Opinion on Drug Discovery·Fabio ModaGiuseppe Legname
Aug 23, 2020·International Journal of Molecular Sciences·Stephanie M Grant, Sharon DeMorrow
Oct 16, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Vladimir I MuronetzMatej Sova
Jan 7, 2022·Cell and Tissue Research·Elisa UliassiGiuseppe Legname

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Methods Mentioned

BETA
electrophoresis
transmission electron microscopy
X-ray
PCR

Software Mentioned

RENAGE
ShIC RENAGE
Origin
ImageJ
ShIC
ProtParam
OriginLab

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